Studies on ovine interleukin-1

Interleukin-1 (IL-1) is a key mediator of infection, inflammation and immunity and two different IL-1 proteins are known to exist, IL-la andIL-lJ3. Each is synthesised as a proprotein, Mr = 31kD, which is subsequent cleaved to yield the mature protein, Mr = 17.5kD. Whereas both forms of IL-la show equivalent biological activities, IL-1 (3 requires cleavage with resultant conformational change for optimal activity. IL-1 has been extensively studied in human and murine systems but at the time this project was initiated, nothing was known about its actions in the sheep and no reagents were available with which to study ovine IL-1. This thesis describes the successful cloning and expression of biologically active ovine IL-la and IL-1 (3 and their use in determining IL-1 receptor (IL-1R) expression by ovine alveolar macrophages (M) and afferent lymph dendritic cells (DC).Lipopolysaccharide (LPS) stimulated Mφ were used as the source of IL-1 mRNA. The specific IL-1 cDNAs were amplified by polymerase chain reaction, cloned into pTZ18R/19R vectors and sequenced. Ovine IL-lα and IL-1ß were found to be 97% and 96% identical to their bovine counterparts and 81% and 78% identical, respectively, to the human sequences. Translation of the nucleotide sequences shows that ovine IL-lα has 97% and 72% identity with bovine and human IL1α respectively and ovine IL-1ß has 95% and 62% identity with bovine and human IL- 1ß respectively. Use of the cloned IL-lcDNAs for northern blot analysis of IL-1 mRNA production by Mφ, showed IL-lα mRNA to reach maximal levels at around 6h and IL-1ß mRNA at around 4h after LPS stimulation.The proprotein (p) and mature protein (m) forms of ovine IL-1α and IL-1ß have been expressed as fusion proteins with yeast p1, using the Ty-vlp system of British Biotechnology Ltd. The fusion protein self-assembles in the form of a virus-like particle (vlp), from which the rIL-1 is cleaved by the action of Factor Xa restriction protease (FXa). FXa cleavage of IL-lap:p1 vlps revealed a relaxed FXa recognition site within IL-1lαp which yielded a putative N-terminus at Ser-120, equivalent to the N-terminus of human EL-lαm. It is proposed that FXa may be one of the natural processing enzymes for IL-1a. The activities of purified IL-1 mature proteins on ovine thymocytes were ~2.5xl0⁷ U/mg EL-1a and ≥1.25xl0⁶ U/mg IL-1ß. Activities on ovine xiphoid cartilage were ten times less. Ovine rIL-1 shows a species preference for homologous thymocytes and cross-species sequence analysis highlights 5 amino acid residues which may be of relevance in this context. Ovine rIL-1ßp is biologically active, but about 5 times less so than IL-1ßm.By using ovine ¹²⁵I-rIL-1, ~16500 IL-1R/cell (Kd ~4.6pM) for IL-1ß and ~2600 IL-lR/cell (Kd ~56pM) for IL-lα were detected on Mφ. ≤24% of Mφ expressed IL-1R. DC, isolated from lymph from cannulated sheep pseudoafferent lymph ducts, expressed ~510 IL-1R/cell (Kd ~30pM) for IL1α and ~350 IL-1R/cell (Kd ~160pM) for IL-1ß. Quantitation of IL-1R expressed by fresh DC has not previously been reported for any species, as far as I am aware. Following secondary ovalbumin challenge of primed sheep, DC expression of receptors for IL-1α, but not for IL-1ß, is greatly upregulated. Two peaks of increased expression were detected, a transient increase at around 4h followed by an increase to maximal expression of ≥21700 IL-1α sites/cell (Kd ~181pM) between 43-72h. Only a very small proportion of individual DC were seen to bind IL-1 even after secondary antigen challenge

Traumebehandling gjennom bilder: En metasyntese av klienters erfaringer med kunstterapi

En betydelig andel av befolkningen opplever potensielt traumatiserende hendelser, som utgjør en trussel mot liv eller helse. For noen vil dette medføre psykologiske og somatiske reaksjoner og symptomer. Traumehendelsen kan ha overskredet personens integrative kapasitet, og sensoriske, emosjonelle og kognitive komponenter kan dermed lagres som løsrevede fragmenter. Emosjonell dysregulering og dissosiasjon kan være vanlige reaksjoner i etterkant. Kunstterapi kan antas å være en relevant behandlingsform for symptomene, blant annet på grunn av traumeminnenes overveldende og ikke-språklige karakter. Dette legger grunnlag for problemstillingen: Hvordan opplever personer med traumeerfaring kunstterapi som behandlingsmetode? Spørsmålet ble belyst gjennom forskningsmetoden kvalitativ metasyntese. Systematisk litteratursøk resulterte i syv inkluderte artikler, hvorav identifiserte hovedtemaer var: 1. Kunstterapi skaper rom til å forholde seg til fortid, og 2. Kunstterapi gir tilgang til nye måter å respondere på. Resultatene viste at et visuelt og eksternt uttrykk kan bidra til gradvis eksponering for traumeerfaringer. I tillegg kan visuelle og metaforiske aspekter ved behandlingen bidra til å tilgjengeliggjøre og uttrykke minnefragmenter som er lagret ikke-verbalt og subkortikalt. En dialektisk tilnærming, bestående av visuelle uttrykk og verbal forståelse, kan utgjøre en prosess der minnefragmenter gradvis integreres. Videre kan det utforskende og skapende aspektet ved kunstterapi, bidra til å fremme ressurser og identitet.A significant proportion of the population experiences potentially traumatizing events that pose a threat to life or health. For some, this will entail both psychological and somatic reactions and symptoms. The traumatic event may have exceeded the person's integrative capacity, where sensory, emotional and cognitive components might be stored as detached fragments. Emotional dysregulation and dissociation may be common symptoms afterwards. Art therapy can be assumed to be a relevant form of treatment for these reactions, partly due to the non-linguistic nature of the traumatic memories. This forms the basis for the problem statement: How do people with trauma exposure experience art therapy as a treatment method? The question was elucidated using the research method qualitative metasynthesis. A systematic literature search resulted in seven included articles, wherein the main themes identified were: 1. Art therapy creates space to be able to relate to the past, and 2. Art therapy provides access to new ways of responding. The results showed that a visual and external expression can contribute to a graduated exposure to trauma experiences. In addition, visual and metaphorical aspects of the treatment may help accessing and expressing fragments of memory that are stored non-verbally and subcortically. A dialectic approach, consisting of visual expression and verbal understanding, may constitute a process in which memory fragments are gradually integrated. Furthermore, the exploratory and creative aspect can help emphasize resources and identity.Hovedoppgave psykologprogrammetPROPSY317PRPSY

Comparison of pre-analytical conditions for quantification of serotonin in platelet-poor plasma

Background: Reported concentrations of serotonin in platelet-poor plasma (PPP) in healthy subjects vary widely due to different pre-analytical procedures. Aim: To examine how different pre-analytical conditions affect the measured concentration of serotonin in PPP. Method: Six pre-analytical protocols were compared for preparation of PPP from EDTA whole blood for quantification of serotonin from nine healthy individuals. Three combinations of centrifugation with a mild centrifugation of gel-free EDTA tubes followed by a stronger centrifugation were compared to single-stage centrifugation of EDTA tubes with separator gel and heat shock treatment of blood prior to centrifugation. All samples were analysed using the same enzyme linked immunosorbent assay (ELISA) method. Results: Findings show that two consecutive centrifugations; first a mild centrifugation at 100 or 200×g followed by centrifugation at 4500 or 14500×g resulted in the lowest serotonin concentration in PPP. Conclusion: Two successive centrifugations to produce PPP for serotonin analysis; first a mild centrifugation to avoid mechanical stress on the platelets, and next a stronger centrifugation to remove platelets, is superior to the use of gel tubes and heat shock treatment.publishedVersio

Sustainable Investments in the Norwegian Stock Market

Master of Science in Business//Siviløkonom - Nord universitet 201

Plasma levels of guanylins are reduced in patients with Crohn’s disease

Background: Guanylin (GN) and uroguanylin (UGN) are endogenous ligands for the intestinal receptor guanylate cyclase C (GC-C), an important regulator of intestinal fluid homeostasis. Gene expression and protein levels of GN are suppressed in inflamed intestinal tissue from patients with inflammatory bowel disease (IBD), but knowledge about plasma levels of guanylins in these conditions is sparse. We aimed to investigate the fasting plasma levels of the prohormones proGN and proUGN in patients with Crohn’s Disease (CD) and relate these to levels found in persons with other diarrheal conditions, as well as persons with normal bowel habits. Methods: Plasma from patients with CD, patients with Familial GUCY2C Diarrheal Disease (FGDS), diarrhea-predominant irritable bowel syndrome (IBS-D) and healthy controls (HC) was analyzed using ELISA assays. Results: Significantly lower fasting plasma levels of proguanylins were found in CD and FGDS patients, compared to HC. In CD patients, plasma proGN levels correlated negatively with Harvey Bradshaw Index and with number of stools/24 h. Conclusion: Our data indicate that diarrhea may be a determinant for levels of proGN in plasma, and should be further explored in studies of different diarrheal disorders.acceptedVersio

The intronic BRCA1 c.5407-25T>A variant causing partly skipping of exon 23—a likely pathogenic variant with reduced penetrance?

Rare sequence variants in the non-coding part of the BRCA genes are often reported as variants of uncertain significance (VUS), which leave patients and doctors in a challenging position. The aim of this study was to determine the pathogenicity of the BRCA1 c.5407-25T>A variant found in 20 families from Norway, France and United States with suspected hereditary breast and ovarian cancer. This was done by combining clinical and family information with allele frequency data, and assessment of the variant’s effect on mRNA splicing. Mean age at breast (n = 12) and ovarian (n = 11) cancer diagnosis in female carriers was 49.9 and 60.4 years, respectively. The mean Manchester score in the 20 families was 16.4. The allele frequency of BRCA1 c.5407-25T>A was 1/64,566 in non-Finnish Europeans (gnomAD database v2.1.1). We found the variant in 1/400 anonymous Norwegian blood donors and 0/784 in-house exomes. Sequencing of patient-derived cDNA from blood, normal breast and ovarian tissue showed that BRCA1 c.5407-25T>A leads to skipping of exon 23, resulting in frameshift and protein truncation: p.(Gly1803GlnfsTer11). Western blot analysis of transiently expressed BRCA1 proteins in HeLa cells showed a reduced amount of the truncated protein compared with wild type. Noteworthily, we found that a small amount of full-length transcript was also generated from the c.5407-25T>A allele, potentially explaining the intermediate cancer burden in families carrying this variant. In summary, our results show that BRCA1 c.5407-25T>A leads to partial skipping of exon 23, and could represent a likely pathogenic variant with reduced penetrance.publishedVersio

Changes in balance ability, power output, and stretch-shortening cycle utilisation after two high-intensity intermittent training protocols in endurance runners

Purpose: This study aimed to describe the acute effects of 2 different high-intensity intermittent trainings (HIITs) on postural control, countermovement jump (CMJ), squat jump (SJ), and stretch-shortening cycle (SSC) utilisation, and to compare the changes induced by both protocols in those variables in endurance runners.Methods: Eighteen recreationally trained endurance runners participated in this study and were tested on 2 occasions: 10 runs of 400 m with 90 s recovery between running bouts (10 × 400 m), and 40 runs of 100 m with 30 s recovery between runs (40 × 100 m). Heart rate was monitored during both HIITs; blood lactate accumulation and rate of perceived exertion were recorded after both protocols. Vertical jump ability (CMJ and SJ) and SSC together with postural control were also controlled during both HIITs.Results: Repeated measures analysis revealed a significant improvement in CMJ and SJ during 10 × 400 m (p < 0.05), whilst no significant changes were observed during 40 × 100 m. Indexes related to SSC did not experience significant changes during any of the protocols. As for postural control, no significant changes were observed in the 40 × 100 m protocol, whilst significant impairments were observed during the 10 × 400 m protocol (p < 0.05).Conclusion: A protocol with a higher number of shorter runs (40 × 100 m) induced different changes in those neuromuscular parameters than those with fewer and longer runs (10 × 400 m). Whereas the 40 × 100 m protocol did not cause any significant changes in vertical jump ability, postural control or SSC utilisation, the 10 × 400 m protocol impaired postural control and caused improvements in vertical jumping tests.Open Access funded by Shanghai University of Spor